甲基营养型芽胞杆菌NKG-1对番茄灰霉病的防治效果及全基因组分析

doi:10.16409/j.cnki.2095-039x.2024.02.024

摘要/Abstract

摘要： 为了研究甲基营养型芽胞杆菌NKG-1对番茄灰霉病的防治效果，采用NKG-1发酵液处理番茄灰霉菌，观察番茄灰霉菌菌丝生长状态，测定菌丝干重、抑菌率，调查其对番茄离体叶片、苗期灰霉病防治效果。结果显示甲基营养型芽胞杆菌NKG-1显著抑制番茄灰霉菌菌丝的生长和产孢，对番茄离体叶片灰霉病防治效果达80.00%，对苗期番茄灰霉病的防治效果达73.14%。通过对NKG-1全基因组进行分析预测，发现该菌含有12个次级代谢产物基因簇，其中脂肽类抗生素表面活性素和丰原素具有抑菌作用，且丰原素已被证实具有强烈的抗真菌活性。本研究为甲基营养型芽胞杆菌NKG-1防治番茄灰霉病的应用和解析防病机理提供了重要的理论基础。

关键词: 甲基营养型芽胞杆菌, 番茄灰霉病, 全基因组, NKG

Abstract: To investigate control effect of Bacillus methylotrophicus NKG-1 on tomato gray mold, dry weight of B. cinereaand inhibition rate and control efficacy of NKG-1 on tomato gray mold were investigated after treatment with the fermentation broth of NKG-1. The results showed that NKG-1 significantly inhibited the hyphae growth and spore production of B. cinerea. The control efficacy of NKG-1 against gray mold on tomato leaves in vitro was 80.00%, and that on tomato plant at seedling stage was 73.14%. By analyzing and predicting the whole genome of NKG-1, it was found that the NKG-1contained 12 secondary metabolite gene clusters. Among them, lipopeptide antibiotics surfactant and fengycin have antibacterial effects. Fengycin has been proved to have strong antifungal activity. The above results provide an important theoretical basis for application of B. methylotrophicus NKG-1 to control tomato gray mold and are helpful to analyze the biological control mechanism of B. methylotrophicus NKG-1.

Key words: Bacillus methylotrophicus, tomato gray mold, whole genome, NKG-1

中图分类号:

S476

张丝雨, 高文静, 张丙翠, 杨吉, 刘焕珊, 刘炳花. 甲基营养型芽胞杆菌NKG-1对番茄灰霉病的防治效果及全基因组分析[J]. 中国生物防治学报, 2024, 40(3): 679-689.

ZHANG Siyu, GAO Wenjing, ZHANG Bingcui, YANG Ji, LIU Huanshan, LIU Binghu. Control Effect of Bacillus methylotrophicus NKG-1 on Tomato Gray and Its Genome-Wide Analysis[J]. Chinese Journal of Biological Control, 2024, 40(3): 679-689.

参考文献

[1] 李宝聚, 陈立芹, 孟伟军, 等. 湿度调控对番茄灰霉病菌侵染的影响[J]. 植物病理学报, 2003, 33(2): 167-169.
[2] 杨利敏, 仝赞华, 郭立华, 等. 番茄灰霉生防菌CQ的分子鉴定及其生防效果研究[J]. 中国生物防治学报, 2015, 31(6): 956-960.
[3] 姚士桐, 陆志杰, 金周浩, 等. 春季大棚番茄灰霉病发生规律及影响因子分析[J]. 中国农学通报, 2011, 27(10) :194-198.
[4] 卜元卿, 孔源, 智勇, 等. 化学农药对环境的污染及其防控对策建议[J]. 中国农业科技导报, 2014, 16(2): 19-25.
[5] 王秋颖. 粉红粘帚霉引发的番茄抗灰霉病的抗病信号传导通路分析[D]. 哈尔滨: 东北农业大学, 2019.
[6] 周蒙. 中国生物农药发展的现实挑战与对策分析[J]. 中国生物防治学报, 2021, 37(1): 184-192.
[7] Compant S, Duffy B, Nowak J,et al. Use of plant growth promoting bacteria for biocontrol of plant diseases: principles, mechanisms of action, and future prospects[J]. Applied and Environmental Microbiology, 2005, 71:4951-4959.
[8] Haidar R, Fermaud M, Calvo-Garrido C,et al. Modes of action for biological control ofBotrytis cinerea by antagonistic bacteria[J]. Phytopathologia Mediterranea, 2016, 55: 13-34.
[9] Lu X H, Jiao X L, Hao J J,et al. Characterization of resistance to multiple fungicides inBotrytis cinerea populations from Asian ginseng in northeastern China[J]. European Journal of Plant Pathology, 2016, 144: 467- 476.
[10] Nicot P C, Stewart A, Bardin M,et al. Biological Control and Biopesticide Suppression of Botrytis-Incited Disease[M]//Fillinger S, Elad Y. Botrytis-the Fungus, the Pathogen and Its Management in Agricultural Systems, Cham: Springer International Publishing, 2016, 165-187.
[11] 司方洁, 任金瑶, 黄涛祥, 等. 贝莱斯芽胞杆菌5YN8生物被膜在防治番茄灰霉病过程中的功能研究[J]. 中国生物防治学报, 2022, 38(5): 1223-1230.
[12] 裴莹莹. 贝莱斯芽胞杆菌WZ-37粗提液分离纯化及对番茄灰霉病防效研究[D]. 哈尔滨: 东北农业大学, 2022.
[13] 郭庆港, 刘高鸽, 陈秀叶, 等. 枯草芽胞杆菌HMB19198菌株抑菌物质的鉴定及其对番茄灰霉病的防治[J]. 植物病理学报, 2022, 52(2): 247-255.
[14] 李凤硕. 两株生防细菌的复合菌剂对番茄灰霉病防效的研究[D]. 哈尔滨: 东北农业大学, 2022.
[15] 邱德文. 生物农药的发展现状与趋势分析[J]. 中国生物防治学报, 2015, 31(5): 679-684.
[16] 周华飞, 罗楚平, 王晓宇, 等. 枯草芽胞杆菌Bs916突变体库的构建和抑制水稻细菌性条斑病菌相关基因的克隆[J]. 中国农业科学, 2013, 11: 2232-2239.
[17] Soledad Figueredo M, Laura Tonelli M, Taurian T,et al. Interrelationships betweenBacillus sp. CHEP5 andBradyrhizobium sp. SEMIA6144 in the induced systemic resistance againstSclerotiumrolfsii and symbiosis on peanut plants[J]. Journal of Biosciences, 2014, 39(5): 877-885.
[18] Raton T O, Giro Z G, Diaz M S,et al. In vitro growth inhibition ofCurvularia gudauskasii byBacillus subtilis[J]. Annals Microbiology, 2012, 62(62): 545-551.
[19] Gong A D, Li H P, Yuan Q S,et al. Antagonistic mechanism of iturin A and plipastatin A fromBacillus amyloliquefaciens S76-3 from wheat spikes againstFusarium graminearum[J]. PLoS ONE, 2015, 10(2): e0116871.
[20] Waewthongrak W, Pisuchpen S, Leelasuphakul W. Effect ofBacillus subtilis and chitosan applications on green mold (Penicilium digitatumSacc.) decay in citrus fruit[J]. Postharvest Biology and Technology, 2015, 99: 44-49.
[21] Scholz R, Vater J, Budiharjo A,et al. Amylocyclicin, a novel circular bacteriocin produced byBacillus amyloliquefaciens FZB42[J]. Journal of Bacteriology, 2014, 196(10): 1842-1852.
[22] Kim P I, Ryu J, Kim Y H,et al. Production of biosurfactant lipopeptides iturin A, fengycin, and surfactin A fromBacillus subtilis CMB32 for control ofColletotrichum gloeosporioides[J]. Journal of Microbiology and Biotechnology, 2010, 20(1): 138-145.
[23] Ge B B, Liu B H, Thinnthinn N,et al.Bacillus methylotrophicus strain NKG-1, isolated from Changbai Mountain，China，has potential applications as a biofertilizer or biocontrol agent[J]. PLoS ONE, 2016, 11(11): e0166079.
[24] 赵文珺, 葛蓓孛, 刘炳花, 等. 甲基营养型芽胞杆菌NKG-1对番茄白粉病的防病促生作用研究[J]. 中国农学通报, 2018, 34(1): 104-109.
[25] 黄海. 解淀粉芽孢杆菌Ba168对番茄灰霉病的防治作用[D]. 咸阳: 西北农林科技大学, 2014.
[26] Yanez-Mendizabal V, Usall J, Vinas L,et al. Potential of a new strain ofBacillus subtilisCPA-8 to control the major postharvest diseases of fruit[J]. Biocontrol Science and Technology, 2011, 21(4): 409-426.
[27] 郭明程, 王晓军, 苍涛, 等. 我国生物源农药发展现状及对策建议[J]. 中国生物防治学报, 2019, 35(5): 755-758.
[28] 马佳, 李颖, 胡栋, 等. 芽胞杆菌生物防治作用机理与应用研究进展[J]. 中国生物防治学报, 2018, 34(4): 639-648.
[29] 谢鑫, 张踞林, 王红宁, 等. 芽孢杆菌中天然脂肽类抗生素的合成及作用机制研究进展[J]. 中国抗生素杂志, 2021, 46(5): 362-370.
[30] Zhao H, Shao D, Jiang C,et al. Biological activity of lipopeptides fromBacillus[J]. Applied Microbiology and Biotechnology, 2017, 101(15): 5951-5960.
[31] Asaka O, Shoda M. Biocontrol ofRhizoctonia solani damping-off of tomato withBacillus subtilis RB14[J]. Applied and Environmental Microbiology, 1996, 62(11): 4081-4085.
[32] Bais HP, Fall R, Vivanco J M. Biocontrol ofBacillus subtilis against infection of Arabidopsis roots byPseudomonas syringae is facilitated by biofilm formation and surfactin production[J]. Plant Physiology, 2004, 134(1): 307-319.
[33] Yanez-Mendizabal V, Zeriouh H, Vinas I,et al. Biological control of peach brown rot (Monilinia spp.) byBacillus subtilis CPA-8 is based on production of fengycin-like lipopeptides[J]. European Journal of Plant Pathology, 2012, 132: 609-619.